| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| TFEB Rabbit mAb Coated Microwells | 68473 | 96 tests |
|
+4C |
| TFEB Mouse Detection mAb | 79965 | 1 ea |
|
+4C |
| HRP Diluent | 13515 | 5.5 ml |
|
+4C |
| TMB Substrate | 7004 | 11 ml |
|
+4C |
| STOP Solution | 7002 | 11 ml |
|
+4C |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml |
|
-20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The rapid protocol (RP) PathScan® RP Total TFEB Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of TFEB protein in a reduced assay time of 1.5 hours. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with TFEB in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of TFEB protein. Learn more about your ELISA kit options here.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
Background
Transcription factor EB (TFEB) is a member of the Myc-related, bHLH leucine-zipper family of transcription factors that drives the expression of a network of genes known as the Coordinated Lysosomal Expression and Regulation (CLEAR) network (1,2). TFEB specifically recognizes and binds regulatory sequences within the CLEAR box (GTCACGTGAC) of lysosomal and autophagy genes, resulting in the upregulated expression of genes involved in lysosome biogenesis and function, and regulation of autophagy (1,2). TFEB is activated in response to nutrient deprivation, stimulating translocation to the nucleus where it forms homo- or heterooligomers with other members of the microphthalmia transcription factor (MiTF) subfamily and resulting in upregulation of autophagosomes and lysosomes (3-5). Recently, it has been shown that TFEB is a component of mammalian target of rapamycin (mTOR) complex 1 (mTORC1), which regulates the phosphorylation and nuclear translocation of TFEB in response to cellular starvation and stress (6-9). During normal growth conditions, TFEB is phosphorylated at Ser211 in an mTORC1-dependent manner. Phosphorylation promotes association of TFEB with 14-3-3 family proteins and retention in the cytosol. Inhibition of mTORC1 results in a loss of TFEB phosphorylation, dissociation of the TFEB/14-3-3 complex, and rapid transport of TFEB to the nucleus where it increases transcription of CLEAR and autophagy genes (10). TFEB has also been shown to be activated in a nutrient-dependent manner by p42 MAP kinase (Erk2). TFEB is phosphorylated at Ser142 by Erk2 in response to nutrient deprivation, resulting in nuclear localization and activation, and indicating that pathways other than mTOR contribute to nutrient sensing via TFEB (3).
- Sardiello, M. et al. (2009) Science 325, 473-7.
- Sardiello, M. and Ballabio, A. (2009) Cell Cycle 8, 4021-2.
- Settembre, C. et al. (2011) Science 332, 1429-33.
- David, R. (2011) Nat Rev Mol Cell Biol 12, 404.
- Cuervo, A.M. (2011) Science 332, 1392-3.
- Peña-Llopis, S. et al. (2011) EMBO J 30, 3242-58.
- Settembre, C. and Ballabio, A. (2011) Autophagy 7, 1379-81.
- Peña-Llopis, S. and Brugarolas, J. (2011) Cell Cycle 10, 3987-8.
- Settembre, C. et al. (2012) EMBO J 31, 1095-108.
- Martina, J.A. et al. (2012) Autophagy 8, 903-14.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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